Abstract：
This study was designed to test the ability of ex vivo antibody-coated intravascular devices to capture genetically engineered pig endothelial colony-forming cells (ECFCs) as proof of concept for their potential for in vivo targeted drug delivery. Human α-calcitonin gene-related peptide (α-CGRP) was chosen as the therapeutic molecule as it is unsuitable for systemic administration due to its potent peripheral arterial vasodilatory effect and short half-life in blood, requiring local delivery to yield therapeutic benefit in a particular vascular bed. H-2Kk, a murine leukocyte surface antigen, served as the selection marker for genetically modified ECFCs. H-2Kk antibody was immobilized on electropolished cobalt-chromium (CC) discs, CC stents and ePTFE grafts through dopamine self-polymerization. The functionalized surface was integral and smooth, lacked or had significantly reduced chemical signals specific for substrates. Pig bone marrow-derived ECFCs transfected with a plasmid constructed for H-2Kk and α-CGRP expression produced H-2Kk on cell surface and biologically active α-CGRP in culture medium. H-2Kk antibody-coated substrates bound H-2Kk ECFCs but not control ECFCs in vitro. Bare or only dopamine-coated substrates did not bind H-2Kk ECFCs. These data suggest that implantation of antibody functionalized devices combined with injection of genetically modified ECFCs could be potentially applied for targeted drug delivery.
摘要：
本研究旨在测试离体抗体包被的血管内装置捕获基因工程猪内皮集落形成细胞（ECFC）的能力，作为其体内靶向药物递送潜力的概念证明。选择人α-降钙素基因相关肽（α-CGRP）作为治疗分子，因为由于其有效的外周动脉血管舒张作用和血液半衰期短，因此不适合全身给药。需要局部递送以在特定血管床中产生治疗益处。H-2Kk,一种鼠白细胞表面抗原,作为遗传修饰的ECFC的选择标记。将H-2Kk抗体固定在电抛光的钴铬（CC）圆盘上，CC支架和ePTFE通过多巴胺自聚合接枝。功能化的表面是完整的和光滑的，缺乏或具有显著降低的底物特异性化学信号。用构建用于H-2Kk和α-CGRP表达的质粒转染的猪骨髓来源的ECFCs在细胞表面产生H-2Kk，并在培养基中产生生物活性的α-CGRP。H-2Kk抗体包被的底物在体外结合H-2KkECFCs但不控制ECFCs。裸露或仅多巴胺包被的底物不结合H-2KkECFC。这些数据表明,抗体功能化装置的植入与遗传修饰的ECFC的注射组合可以潜在地应用于靶向药物递送。
